[Analysis of CgGPD gene promoter from Candida glycerinogenes by fluorescent protein].
OBJECTIVE; We cloned the promoter of glycerol-3-phosphate dehydrogenase gene(CgGPD) from the Candida glycerinogenes, and studied its functional regulation under high osmotic stress condition.


METHODS
We amplified the 950 bp promoter of CgGPD from C. glycerinogenes and the green fluorescent protein gene (gfp) from pCAMBIA1302 vector by PCR and introduced them into a modified vector pYX212-zeocin simultaneously. The recombinant plasmid pYX212-zeocin harboring both the promoter of CgGPD and gene gfp was transformed into S. cerevisiae W303-1A by electroporation. In the medium containing glucose with different concentrations for culturing the recombinant strain S. cerevisiae W303-1A-GFP the green fluorescence was detected by fluorescent microscopy.


RESULTS
The gene gfp was functionally expressed under the control of the promoter of CgGPD in S. cerevisiae. Furthermore, the expression of the gene gfp at different level was conducted by the different osmotic stress for the recombinant strain. The green fluorescence was less intensive when the concentration of glucose was low for culturing the recombinant strain, but it became much more intensive when the concentration of glucose increased.


CONCLUSIONS
The promoter of CgGPD is an inducible promoter that can be induced significantly by the high concentration of glucose. The promoter will facilitate further studies on the mechanism of glycerol synthesis from C. glycerinogenes WL2002-5 under osmotic stress conditions.
Ding
Chunsheng
C
Key Laboratory of Industrial Biotechnology, Ministry of Education, Research Center of Industrial Microbiology, Jiangnan University, Wuxi 214122, China. ding810323@163.com
Rao
Zhiming
Z
Zhuge
Bin
B
Shen
Wei
W
Chen
Xianzhong
X
Fang
Huiying
H
Zhuge
Jian
J
D002175:Candida / Q000235:genetics*; D002237:Carbohydrate Dehydrogenases / Q000235:genetics*; D003001:Cloning, Molecular; D005453:Fluorescence; D005822:Genetic Vectors / Q000235:genetics; D005990:Glycerol / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014170:Transformation, Genetic
D049452:Green Fluorescent Proteins; D002237:Carbohydrate Dehydrogenases; C056470:3-glycerophosphate dehydrogenase; D005990:Glycerol